Analysis of protein extracts from films and gloves produced with hevea and guayule latex

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Analysis of protein extracts from

films and gloves produced with

hevea and guayule latex

M Daniau, G Olivier, A Souverain, D Pioch, S Palu, M Rossignol, S Hem, D Sauvaire, C Mourton-Gilles Biorubber for Europe in Global Perspective, Wageningen, September 25th, 2012

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Scientific

context

Objective

Collaborative study between CIRAD and National Health product Agency To compare protein content of Guayule and Hevea device extracts

Latex allergy is due to the présence of hevea proteins in rubber goods

(cleaning gloves, nipples, toys,…) and medical devices (gloves, catheter, syringe,…).

Latex allergy is a real health problem

Up to 17% of health care workers suffer from type I hevea allergy Up to 16% of intra-operative shock are due to this allergy

Presence of cross allergy with some fruits (banana, kiwi,…) A solution to this problem could be to develop devices

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Agence nationale de sécurité du médicament et des produits de santé

361.76 3.13 26.43 5.35 66.53 12.46 431.22 0 50 100 150 200 250 300 350 400 450 Protein quantity (µg/g)

Hevea f ilms Guayule f ilms Powered f ree lat ex gloves

St erile powered f ree lat ex gloves

Hevea lat ex gloves (CIRAD/ CTTM ) Guayule lat ex Yulex gloves (CIRAD/ CTTM ) Guayule lat ex EU-PEARLS gloves (CIRAD/ CTTM )

Determination of total protein content

in films and gloves extracts

Protein quantity in guayule films are 115 times lower than in hevea films and 9 times than in medical gloves

Gloves made with commercial guayule latex contain less protein than hevea prototype gloves.

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1.16E-02 4.30E-04 2.52E-04 0 6.33E-04 7.28E-04 0 0.00E+00 2.00E-03 4.00E-03 6.00E-03 8.00E-03 1.00E-02 1.20E-02 Hev b1 (µg/g)

Hevea film Guayule film Pow ered free latex gloves

Steril pow ered free latex gloves Hevea latex gloves (CIRAD/CTTM) Guayule latex Yulex gloves (CIRAD/CTTM) Guayule latex EU-PEARLS gloves (CIRAD/CTTM) 2.17E-01

1.87E-04 9.36E-03 ₀ 7.22E-03 2.15E-03 0 0.00E+00 5.00E-02 1.00E-01 1.50E-01 2.00E-01 2.50E-01 Hev b3 (µg/g)

Steril pow ered free latex gloves Hevea latex gloves (CIRAD/CTTM) Guayule latex Yulex gloves (CIRAD/CTTM) Guayule latex EU-PEARLS gloves (CIRAD/CTTM)

Determination of allergen content

in films and gloves extracts

Allergen Hevb3 is present in the hevea films and in small amount in medical gloves and guayule gloves made with

commercial latex

Allergen Hevb1 is present in the hevea films and in small amount in guayule extracts

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0 2.33E-05 4.68E-03 6.71E-05 8.31E-04 0 0 0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03 5.0E-03 Hev b5 (µg/g)

Steril pow ered free latex gloves

Hevea latex gloves (CIRAD/CTTM) Guayule latex Yulex gloves (CIRAD/CTTM)

Guayule latex EU-PEARLS gloves (CIRAD/CTTM) 1.02E-02 1.86E-04 2.07E-03 2.10E-04 5.50E-03 2.27E-03 1.46E-03 0.00E+00 2.00E-03 4.00E-03 6.00E-03 8.00E-03 1.00E-02 1.20E-02 Hev b6.02 (µg/g)

Steril pow ered free latex gloves

Hevea latex gloves (CIRAD/CTTM) Guayule latex Yulex gloves (CIRAD/CTTM)

Guayule latex EU-PEARLS gloves

(CIRAD/CTTM)

Determination of allergen content

in films and gloves extracts

Allergen Hevb6.02 is present in the hevea films and gloves but also in

guayule films and gloves

Allergen Hevb5 is present in the hevea gloves

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Using WB and labelling with anti-hevea Mabs (Hevb1,Hevb3,Hevb5 and Hevb6.02)

Using LC ESI MS/MS mass spectrometry: collaboration with INRA

Some hevea allergens were identified

Rubber elongation factor

Hevb1

Glucan endo1,3 beta glucosidase:

Hev b2

Hevamin A :

Hev b14

Pro-Hevein:

Hev b6.01

One guayule protein was identified

Allene oxyde synthase

Identification of proteins in guayule films extracts

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Conclusion

Protein quantity in guayule films is about 100 times lower than in hevea films Only Hevb1 and Hevb6.02 were detected in guayule films using Fit Kits

Gloves made with commercial guayule latex contain less protein than hevea gloves But, gloves made with CIRAD guayule latex contain large protein amounts

Small amounts of Hevb1, Hevb3 and Hevb6.02 were detected in guayule glove prototypes

Using mass spectrometry, four hevea allergens were found in guayule film extract Hevb1, Hevb2, Hevb14 and Hev b6.01

We detected Hevb6.02 (hevein) by ELISA or Hevb6.01 (pro-hevein) by MS

Identification of a guayule specific protein (Parthenium argentatum) : Allene oxyde synthase

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